盐酸甲氟喹促进DNA修复减轻辐射诱导的肺上皮细胞损伤OA
Mefloquine HCl promotes DNA repair and alleviates radiation-induced lung epithelial cell injury
目的 探讨盐酸甲氟喹(MQ)对X射线照射所致肺上皮细胞DNA损伤的保护作用.方法 实验将人正常肺上皮细胞(BEAS-2B)分为空白对照组、空白对照+MQ组、单纯照射组、照射+MQ处理组;利用CCK-8实验、EdU-488实验、细胞克隆形成实验观察MQ对X射线照射后细胞增殖和放射敏感性的影响;通过流式细胞术检测不同条件处理BEAS-2B细胞的凋亡及细胞周期情况;通过细胞免疫荧光实验、彗星电泳观察MQ对BEAS-2B细胞DNA双链断裂(DSB)损伤修复的调控功能;利用Western blotting、qPCR和荧光素酶实验研究MQ调控DSB损伤修复的作用机制.结果 CCK-8结果显示MQ 0～40 μmol/L范围内,0～10 μmol/L对BEAS-2B细胞活力无明显抑制作用.与单纯照射组相比,10 Gy照射联合不同浓度(1～10 μmol/L)处理均能增强细胞活力,且10 μmol/L时细胞活力最高,故被选取为后续实验浓度;MQ处理能提高照射后BEAS-2B细胞增殖能力(P<0.05)和克隆形成率(P<0.05),降低辐射敏感性;同时单纯照射组细胞凋亡率为(17.43±0.51)%,而照射+MQ处理组细胞凋亡率降低至(14.03±0.45)%(t=8.621,P<0.05);细胞周期检测发现,单纯照射导致G2/M期阻滞,MQ可缓解由4Gy照射导致的细胞G2/M期阻滞(P<0.05);彗星实验和免疫荧光实验显示,与空白对照组相比,空白对照+MQ处理组的彗星尾矩与γH2AX foci差异无统计学意义;而照射+MQ处理组的彗星尾矩(P<0.05)和γH2AX foci形成数量(P<0.01)均较单纯照射组降低,DSB修复效率提高;Western blotting 实验与qPCR实验显示,与空白对照组相比,MQ处理BEAS-2B细胞48 h后,能够促进CtIP启动子活性(P<0.05),从而在mRNA和蛋白水平上上调CtIP的表达.结论 MQ通过上调CtIP表达,促进BEAS-2B的DSB修复能力,进而减轻辐射诱导的肺上皮细胞损伤.
Objective To investigate the protective effect of mefloquine HCl(MQ)against X-ray irradiation-induced DNA damage in lung epithelial cells.Methods Human lung epithelial cells(BEAS-2B)were divided into blank control group,irradiation group,and irradiation+MQ treatment group.The effects of MQ on cell proliferation and radiosensitivity after X-ray irradiation were assessed using CCK-8 assay,EdU-488 assay,and colony formation assay.Apoptosis and cell cycle distribution of BEAS-2B cells with different treatments were detected by flow cytometry.The effect of MQ in promoting DNA double-strand break(DSB)repair was observed using immunofluorescence staining and comet assay,and the molecule ar mechanism was explored using Western blotting,qPCR,and luciferase reporter assays.Results MQ at 0-10 μmol/L did not significantly affect BEAS-2B cell viability.Compared to the irradiated cells,treatment with 0-10 μmol/L MQ enhanced the cell viability,and the effect was the most conspicuous at 10 μmol/L.MQ treatment obviously promoted proliferation and increased clonogenic survival rate of irradiated BEAS-2B cells while reducing their radiosensitivity,and significantly lowered cell apoptosis rate following the irradiation.Cell cycle analysis revealed that MQ alleviated G2/M phase arrest induced by irradiation,and comet assay and immunofluorescence staining showed reduced comet tail moment and γH2AX foci formation in irradiation+MQ group.Western blotting and qPCR demonstrated that MQ treatment for 48 h significantly increased CtIP promoter activity and upregulated CtIP expressions at both the mRNA and protein levels.Conclusion MQ promotes DSB repair in BEAS-2B cells by upregulating CtIP expression and may thus alleviate radiation-induced lung epithelial cell injury.
张燕;温庆秋;黄海波;周美娟;王建宇
广州市第十二人民医院职业环境与健康重点实验室,广东 广州 510620南方医科大学公共卫生学院放射医学系//广东省热带病研究重点实验室,广东 广州 510515广州市第十二人民医院职业环境与健康重点实验室,广东 广州 510620南方医科大学公共卫生学院放射医学系//广东省热带病研究重点实验室,广东 广州 510515广州市第十二人民医院职业环境与健康重点实验室,广东 广州 510620
盐酸甲氟喹放射性肺损伤DNA损伤DNA双链断裂修复
mefloquine HClradiation-induced lung injuryDNA damageDNA double-strand break repair
《南方医科大学学报》 2026 (3)
497-504,8
国家自然科学基金(81903256)广州市市校(院)企联合资助专题(广州市职业环境与健康重点实验室)(2024A03J0434)Supported by National Natural Science Foundation of China(81903256).
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